Exploring a multiplex DNA methylation-based SNP typing method for body fluids identification: As a preliminary report

In forensic investigation, identification of the cellular origin from body fluid can be essential in the crime scene reconstruction. Recently, DNA methylation could potentially be used as a novel marker for body fluid identification. The simultaneous analysis of CpGs and neighboring single nucleotide polymorphisms (SNPs) has been proposed as an efficient assay for body fluids identification. In this study, a multiplex DNA methylation-based SNP typing system was developed. The specificity, sensitivity and detectability in mixtures and degraded samples were explored in our study. As results, four DNA methylation-based semen-specific SNP (SE1-4) showed good specificity, but two markers associative with saliva (SA1) and vaginal fluid (VA3) was observed cross-reactivity sporadically. Interesting, VA3 were found only presented in the female which may be useful for sexual identification. Moreover, this multiplex system successfully amplification in mixtures and aged samples which proves it be used as a valuable protocol in the identification of actual forensic samples. The strategy indicated that the approach was suitable and reliable for the body fluids analysis in mix stains in Han Chinese for forensic purposes.